Regulation of embryonic patterning and adult stem cells of oral appendages

描述（由申请人提供）：了解口腔外胚层附属物如牙齿、味觉乳头和丝状乳头的发育、模式和出生后更新的分子和细胞机制，并确定这些器官中的干细胞和祖细胞群，对于制定再生策略以替代先天性缺失或因疾病而缺失的牙齿至关重要；了解和治疗味觉障碍，包括放射治疗和小分子抗癌药物引起的味觉障碍；以及描述口腔癌中可能失调的增殖控制。Wnt/ -catenin信号通路在许多发育过程中是必需的，在成体干细胞群体的增殖和自我更新中起着关键作用。小鼠的遗传学研究揭示了Wnt/ -catenin信号在牙齿和味觉乳头形态发生的早期阶段的关键功能。在牙齿和味觉乳头发育开始之前，信号被广泛激活，并逐渐局限于牙齿和味觉前体发育的部位，确保了牙齿和味觉器官的正确定位。根据我们的初步数据，我们假设Wnt信号的适当定位需要Wnt配体和分泌的Wnt抑制剂的竞争活性，并且这些通过空间控制口腔上皮细胞的命运、运动和增殖来直接器官形成。

DESCRIPTION (provided by applicant): Understanding the molecular and cellular mechanisms regulating the development, patterning and postnatal renewal of oral ectodermal appendages such as teeth, taste papillae and filiform papillae, and identifying stem and progenitor cell populations in these organs, is critical for developing regenerative strategies to replace missing teeth in cases of congenital absence or loss through disease; for understanding and treating disorders of taste, including those resulting from radiation therapy and small molecule anti-cancer drugs; and for delineating proliferation controls that may be dysregulated in oral cancers. The Wnt/ -catenin signaling pathway is necessary for many developmental processes and plays critical roles in the proliferation and self- renewal of adult stem cell populations. Genetic studies in mice reveal key functions for Wnt/ -catenin signaling at early stages of tooth and taste papilla morphogenesis. Signaling is activated broadly prior to the initiation of tooth and taste papilla development and gradually becomes restricted to sites of tooth and taste precursor development, ensuring correct positioning of tooth and taste organs. Based on our preliminary data, we hypothesize that proper localization of Wnt signaling requires competing activities of Wnt ligands and secreted Wnt inhibitors, and that these direct organ formation by spatially controlling the fates, movements and proliferation of oral epithelial cells. We will use live imaging of embryonic oral explants from mice that express chromatin-localized GFP specifically in Wnt-activated cells or in all basal epithelial cells, together with ubiquitous expression of membrane Tomato, to ask whether genetic deletion of Wnt inhibitors, or loss of a Wnt ligand important for tooth development, cause altered cell movements and/or patterns of proliferation. Filiform and taste papillae of the tongue are continuously renewed in adult life, an a subset of human patients with mutations in WNT10A presents with adolescent onset of oral ectodermal defects including "smooth tongues". We hypothesize that WNT10A/ -catenin signaling controls proliferation and/or survival of adult stem and progenitor cells required for renewal of taste and filiform papillae. To test this we will ask whether loss of Wnt10a in mice affects -catenin signaling and the proliferation, survival, or differentiation of tongue papilla progenitor cells and will fate map Wnt responsive cells in the adult tongue to test whether they include self- renewing progenitors. To determine whether Wnt signaling is necessary for survival of functional progenitors, we will test whether inhibition of filiform and taste papilla proliferaton upon inducible transgenic expression of the Wnt inhibitor DKK1 is reversible after removal of the inducing agent. These experiments will delineate mechanisms controlling proliferation and organ renewal in the oral cavity and will provide important information for designing regenerative strategies.